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Additional file 5: Table S5 Primers utilized in the present study.

Additional file 2: Table S2. Assessment of consanguinity based on PI_HAT values calculated by PLINK.

Additional file 3: Table S3. Methylation values of 855 probes (CpGs) on differentially methylated regions, measured by Infinium MethylationEPIC Kit.

Additional file 1: Table S1. Methylation indices (%) for CpGs determined by pyrosequencing analysis for disease-related iDMRs.

Additional file 7: Figure S2. Violin plots of the β-values for autosomal probes in patient 1, the parents, and 24 control subjects (average).

Additional file 4: Table S4. The list of abnormally methylated 24 probes (CpGs) on non-iDMRs.

Additional file 8: Figure S3. Physical map of the chromosome 14q32.2 imprinted region and that of the magnified MEG3/DLK1:IG-DMR and MEG3:TSS-DMR.

Additional file 6: Figure S1. Exclusion of a heterozygous deletion involving ZNF445 in patient 1.

Table S1. Primers utilized in this study. Table S2. Methylation levels (β-values) of each CpG site in leukocyte DNA samples. Table S3. The results of micosatellite analysis. Figure S1. Methylation analyses of the H19/IGF2:IG-DMR, MEG3:TSS-DMR, and GNAS A/B:TSS-DMR, using leukocyte gDNA samples. Figure S2. Lack of UPD (14) mat and microdeletion in this patient. Figure S3. Representative data in patients with both hypomethylation-type and hypermethylation-type of MLID in the absence of a mutation in causative or candidate genes for MLID. (PDF 1007 kb)

Mohawk homeobox (MKX) has been demonstrated as a tendon/ligament specific transcription factor. The aim of this study was to investigate the role of MKX in ligament/tenogenic differentiation of bone marrow derived mesenchymal stem cells (BMMSCs). Human BMMSCs were treated with 50 ng/ml BMP-12 or transduced with MKX or scleraxis (SCX) adenoviral vector. Gene expression analysis was performed by quantitative reverse transcribed polymerase chain reaction (qRT-PCR). Rat BMMSCs were seeded in a collagen scaffold and transplanted into a rat Achilles tendon defect model. Tenogenesis related gene expressions and histological features were analyzed. BMP-12 induced tenogenesis in BMMSCs as indicated by increased COL1a1, TNXB, DCN and SCX mRNA, and MKX expression increased simultaneously. Rat BMMSCs enhanced defect repair and were still detectable 3 weeks after transplantation. Increased expressions of COL1a1, TNC and TNMD in vivo were also correlated with upregulated MKX. Adenoviral MKX promoted expression of COL1a1, TNXB, and TNMD in BMMSCs. This study demonstrated that MKX gene expression is enhanced during the tenogenic differentiation of BMMSCs in vitro and in vivo, and the adenoviral overexpression of MKX increases tendon extracellular matrix gene expression and protein production. Thus, MKX is a key factor for tenogenic differentiation of MSCs.